血红素氧合酶-1基因转染对大鼠供肝缺血再灌注损伤的抗凋亡作用  

Human heme oxgygenase-1 gene transfer protects rat grafts from IRI by antiapoptotic effect followed by OLT

作　　者：胡明政[1] 吴建武[2] 张伯[2] 秦磊[2] 钱海鑫[2] 王学浩[1] 

机构地区：[1]南京医科大学第一附属医院肝脏外科,210029 [2]苏州大学第一附属医院普外科

出　　处：《中华实验外科杂志》2008年第11期1410-1411,共2页Chinese Journal of Experimental Surgery

基　　金：江苏省临床免疫重点实验室基金资助项目;江苏省自然科学基金资助项目（BK2007057）.

摘　　要：目的应用转染血红素氧合酶-1（HO-1）基因研究其在大鼠供肝缺血再灌损伤（IRI）中的抗凋亡作用。方法将转染HO—1基因的腺病毒载体和空载体分别注入供体大鼠腹腔（n=8），36h后取肝冷保存4h行大鼠原位肝移植。缺血再灌注6h检测肝功能、肝细胞凋亡率和肝组织HO-1、bcl-2、bcl—xl和Caspase-3的表达。结果（1）实验组的肝功能明显改善、肝细胞凋亡率显著低于对照组[（1．09±0．28）％比（8．30±1．08）％，P〈0．01]。（2）Western blot检测肝组织HO-1、bcl-2和bcl—xl，灰度比值实验组显著高于对照组（HO-1：0．275±0．065比0．035±0．03；bcl-2：0，275±0．025比0．06±0．07；bcl—xl：（0．099±0．041比0．064±0．064，P〈0．01），而Caspase-3则显著低于对照组（0．08±0．04比0．21±0．09，P〈0．01）。结论HO-1在供肝IRI中通过上调bcl-2、bcl-xl和下调Caspase-3对供肝有显著的抗凋亡保护作用。Objective To investigate the antiapoptopic effect of heine oxgygenase-1 （ HO-1 ） gene transfer in a rat model of IRI followed by orthotopic liver transplantation （OLT）. Methods Control and experimental donors （ n = 8 ） were intraperitoneally pretreated with Ad-EGFP and Ad-HO-1 respectively 36 h before their livers were harvested. Six h after reperfusion, serum ALT, AST and LDH levels were determined and the apoptotic ratio was analyzed by flow cytometer. The expression of HO-1, bcl-2, bcl-xl and Caspase-3 was detected by Western-blot. Results （ 1 ） The expression level of HO-1 in the experimental group was significantly higher than in the control group （ 0. 275 ± 0. 065 vs 0. 035 ± 0.03, P 〈 0.01 ）. The liver function index was ameliorated and the apoptotic ratio of hepatic cells decreased significantly [ （8.30±1.08） % vs （ 1.09 ±0.28） % ] in the experimental group as compared with those in the control group; （2） The expression of bcl-2 and bel-xl in the experimental group was increased significantly （ bel-2：0.06 ±0.07 vs 0. 275± 0. 025 ; bel-xl：0. 064 ± 0. 064 vs 0. 099 ± 0. 041 ,P 〈 0.01 ） , and the expression of easepase-3 was weakened obviously （0.21 ± 0.09 vs 0.08 ± 0.04,P 〈 0.01 ） as compared with those in the control group. Conclusion HO-1 can alleviate IRI in rat liver by suppressing apoptosis whieh relates to significantly modulating proapoptotie （ Caspase-3 ） and antiapoptotie （ bel-2 and bcl-xl） pathways.

关 键 词：肝 血红素氧合酶-1 缺血 再灌注损伤 

分 类 号：R285.5[医药卫生—中药学]